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The effect of antioxidants and the NFkB p65 pathway in inflammation

Tue, 09/20/2016 - 13:55

NFkB is a transcription factor that plays a role in the expression of genes involved in immune response, inflammation, metastasis, cell survival and more. RelA (p65) is one member of the NFkB mammalian family, alongside other subunits. NFkB subunits have recognition sites on the “Rel” homology domain, where they form protein complexes to bind DNA and regulate gene expression. The RelA (p65) subunit also has an extended carboxy terminal that can act as a transactivator (a form of gene regulation that increases the rate of gene expression in response to stimuli). The NFkB p65 subunit is thought to shuttle between the cytoplasm and the nucleus, however it is also shown to stay localized to the cytoplasm by its inhibitor, IKK beta. Over the past ten years or so, introduction of antioxidants to inflammation or aging has had interesting response on a cellular level. Given that the NFkB pathway regulates the levels of endogenous reactive oxygen species (ROS), it is the natural next step to investigate the role that antioxidant introduction would have on these processes.

Rasheed et al used a NFkB p65 antibody in their research to investigate whether green tea polyphenol epigallocatechin-3-gallate (EGCG) inhibits advanced glycation of tumor necrosis factor alpha. The overarching disease that this pathway relates to is osteoarthritis, however the manner in which age related accumulation of glycation end products activate inflammation is not well understood. Using a NFkB p65 antibody they tested the outcome of inhibition of NFkB activation with the introduction of EGCG to potentially mediate its inhibitory effects. Their first results demonstrated that EGCG inhibited the degradation of IKK-alpha and subsequent nuclear translocation of NFkB p65 using a NFkB p65 antibody in western blot. In addition, they employed an ELISA to take a closer look at whether EGCG could successfully inhibit IKK beta kinase activity. The results of the ELISA showed that indeed IKK beta kinase was inhibited in the presence of EGCG.

Next, a NFkB p65 antibody was used in a study aimed at understanding the potential role of flavonoids, a plant pigment and one of the world’s largest nutrient families, in response to the NFkB pathway. Flavonoids have exhibited natural antioxidant power, as diets rich in flavonoids have shown protection from cancer, neurodegeneration, and cardiovascular disease. Overall, their research proved that Swiss albino mice with a high cholesterol diet, and a high cholesterol diet alongside quercetin (a common flavonoid) had interesting results for inflammation. Using a NFkB p65 antibody in western blot, they found that fat accumulation and weight gain caused a rise in pro inflammatory cytokines and high expression of NFkB. This increase was reversed once the dietary effects were reduced.